目的:利用本实验室已获得的与肺癌特异性结合的十二肽ZS-1(EHMALTYPFRPP)从人类肺癌cDNA文库中筛选出肺癌相关标志物,并进行初步确认,为肺癌的临床早期诊断和靶向治疗奠定基础。方法:本实验以ZS-1为探针,通过将偶联生物素的多肽ZS-1固化在亲和素处理的酶标板,筛选人类肺癌cDNA噬菌体展示文库,经过三轮减性筛选后,挑取与ZS-1特异性结合的噬菌体单克隆、扩增、纯化,再通过PCR技术扩增、产物纯化后,进行DNA测序,测序结果经过生物信息学分析,随后采用组织芯片技术对相关生物大分子进行了检测。结果:通过生物信息学分析首次发现与ZS-1结合的肺癌细胞膜受体蛋白为KIAA1199,随后以KIAA199抗体检测人肺癌组织芯片证实,KIAA1199的阳性检出率为32.8%。结论:从人类肺癌cDNA噬菌体展示文库中筛选出潜在的肺癌肿瘤标志物KIAA1199,为肺癌的早期诊治奠定基础。
AIM:To select a potential biomarker for early lung cancer diagnosis and targeted therapy by using the cancer specific bounded peptide ZS-1 which had already been obtained from the laboratory. METHODS:The peptide ZS-1 marked by biotin was used as a probe to pan out the human lung cancer cDNA phage display library,after 3 rounds of subtraction panning,the specific binding clones of ZS-1 were identified.After amplification and purification,then those DNA sequences were identified and analyzed with bioinformatics.Tissue microarray technology was used to detect biomelecule in tissues.RESULTS:The lung cancer membrane receptor protein KIAA1199 binding ZS-1 was first found by bioinformatics analysis.Subsequently,a specific human lung cancer tissue microarray was detected and analysed by KIAA199 antibody.The results displayed KIAA1199 positive detection rate was 32.8%.CONCLUSION:A tumor biomarker KIAA1199 selected from human lung cancer cDNA library provides a potential tool for early lung cancer diagnosis and therapy.